The F-actin assembling was assessed as previously described (Ge et al., 2016 (link)), samples were incubated in Alexa Fluor 488 conjugated phalloidin reagents (Life Technologies, Waltham, MA, United States) at room temperature for 30 min. Images were visualized with a confocal microscope (Carl Zeiss, Weimar, Germany) and measured using Zen 2011 software (Carl Zeiss, Weimar, Germany).
Alexa fluor 488 conjugated phalloidin reagents
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 488 conjugated phalloidin reagents are fluorescent probes used to detect and visualize actin filaments (F-actin) in cells. The Alexa Fluor 488 dye is covalently attached to phalloidin, a toxin that binds specifically to F-actin. This allows the actin cytoskeleton to be imaged and analyzed using fluorescence microscopy techniques.
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Lab products found in correlation
3 protocols using alexa fluor 488 conjugated phalloidin reagents
1
Visualizing F-actin Cytoskeleton Assembly
2
Fluorescence microscopy of F-actin
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The status of F-actin assembly in each group was determined as described previously (25 (link)). In brief, the samples from each group were incubated in 4% paraformaldehyde (PFA) for 10 min at room temperature and washed with PBS three times. Subsequently, the samples were immersed in Alexa Fluor 488 conjugated phalloidin reagents (Life Technologies, Waltham, MA, USA) at room temperature for 30 min. After being mounted on glass slides, the images were captured with a fluorescence microscope (Carl Zeiss, Weimar, Germany) and analyzed by ImageJ software (ImageJ 1.8, NIH, USA).
3
Actin Filament Polymerization Visualization
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Actin filament polymerization was detected as previously described [2 (link), 3 (link)]. Briefly, neurospheres were incubated in 4% paraformaldehyde for 10 min at room temperature and then washed with PBS three times. Thereafter, the samples were incubated in Alexa Fluor 488-conjugated phalloidin reagents (Life Technologies, Waltham, MA, USA) at room temperature for 30 minutes. After mounting onto glass slides, images were visualized with a confocal microscope (Carl Zeiss, LSM780, Weimar, Germany) and measured using Zen 2011 software (Carl Zeiss, Weimar, Germany).
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